The QIAamp UCP DNA Micro Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Minimal background compared with other suppliers.

DNA was prepared using the QIAamp UCP DNA Micro Kit and kits from other suppliers. The 16S gene was amplified using QuantiTect SYBR Green PCR Master Mix treated with propidium monoazide (PMA) to minimize DNA background from the PCR step. The QIAamp UCP DNA Micro Kit’s proprietary decontamination process ensures minimal background compared with kits from other suppliers. 30 columns from each supplier were tested and contamination was defined as >62.5 16S copies (relative to standard curve) in the eluate.

Performance of ultraclean DNA in NGS.

DNA was prepared using kits from various suppliers, as per manufacturer’s instructions and 100 copies of two different bacterial DNAs (Escherchia coli and Bacillus subtilis) were spiked into the eluates to allow quantification of the bacterial sequencing reads. The 16S gene was amplified using the QuantiTect Probe PCR Master Mix (QIAGEN). A library was constructed and sequenced in triplicates using next-generation sequencing on the Illumina MiSeq. The QIAamp UCP DNA Micro Kit had fewer microbial background reads compared with kits from other suppliers. A Bacterial reads as a percentage of total reads. Spiked bacterial reads are shown in dark blue, other (background) reads in light blue. B Further analysis of the microbial community at the family level, excluding spike. Results shown as a percentage of total reads.

Comparable yields.

Yields obtained from 10 µl whole blood using the QIAamp UCP DNA Micro Kit and standard (non-ultraclean) sample prep kits from various suppliers. The yields were comparable between all four methods.

The QIAamp UCP DNA Micro procedure.

Performance

The QIAamp UCP DNA Micro Kit is suitable for a wide range of sample materials, such as small volumes of blood, animal and human cells, swabs and small tissue samples.

The QIAamp UCP DNA Micro Kit is designed for purification of ultraclean genomic, mitochondrial and microbial DNA from small sample volumes. The kit combines the selective binding properties of a silica-based membrane with elution volumes of between 20 and 100 µl.

DNA yield and purity obtained with the QIAamp UCP DNA Micro Kit are comparable to known methods, but with very little background of foreign DNA (see figure “Comparable yields”).

Samples are lysed under highly denaturing conditions at elevated temperatures in the presence of proteinase K and Buffer AUT. DNA is then bound to the QIAamp UCP MinElute column membrane and two wash steps efficiently wash away contaminants. DNA is eluted from the QIAamp UCP MinElute column using a small volume of Buffer AUE or ultraclean PCR water. Since the elution volume is small, the eluted DNA will be concentrated.

Applications

DNA purified using the QIAamp UCP DNA Micro Kit is suitable for use in NGS and other downstream applications requiring the use of ultraclean DNA.